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Immunomagnetic positive selection of mouse CD138+ plasma cells and plasmablasts from single-cell suspensions of mouse splenocytes or bone marrow, using particle release technology
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Easily isolate highly purified and magnetic particle-free CD138+ plasma cells and plasmablasts from single-cell suspensions of mouse splenocytes, bone marrow, or other tissue samples, using immunomagnetic positive selection, with the EasySep? Release Mouse CD138 Positive Selection Kit. Widely used in published research for more than 20 years, EasySep? combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.
This EasySep? positive selection procedure involves labeling CD138+ cells with antibody complexes and EasySep? Releasable RapidSpheres?. Unlike traditional magnetic particles that stay bound to the target cells, RapidSpheres? have a releasable feature. Desired cells are first labeled with antibodies and these specialized magnetic particles, before being separated, without columns, using an EasySep? magnet. Unwanted cells are simply poured off, while desired cells remain in the tube. Then, bound magnetic particles are removed from the EasySep?-isolated, CD138+ cells using a release agent. Following magnetic cell isolation with this EasySep? Release kit in as little as 37 minutes, the desired cells are immediately available for downstream applications such as flow cytometry, cell culture, or hybridoma generation. Antibody complexes remain bound to the surface of the desired cells and may interact with Brilliant Violet? antibody conjugates, polyethylene glycol-modified proteins, or other chemically related ligands. CD138+ cells isolated from immunized mice are up to 95% pure and can be used with hybridoma generation protocols such as electrofusion.
Learn more about how immunomagnetic EasySep? technology works or how to fully automate immunomagnetic cell isolation with RoboSep?. Explore additional products optimized for your workflow, including those for culture media, supplements, antibodies, and more.
(A) Starting with na?ve C57BL/6 mouse splenocytes, the plasma cell (CD138+CD267 (TACI)+) content of the isolated fraction is typically 77.2 ± 2.4% (mean ± SD using the purple EasySep? Magnet). In the above example, the purities of the start and isolated fractions are 0.6% and 77.6%, respectively.
(B) Starting with immunized BALB/c mouse splenocytes, the plasma cell (CD138+CD267 (TACI)+) content of the isolated fraction is typically 86.1 ± 7.4% (mean ± SD using the purple EasySep? Magnet). In the above example, the purities of the start and isolated fractions are 1.2% and 91.6%, respectively.
C) Starting with na?ve C57BL/6 mouse bone marrow, the plasma cell (CD138+CD267 (TACI)+) content of the isolated fraction is typically 62.0 ± 13.1% (mean ± SD using the purple EasySep? Magnet). In the above example, the purities of the start and isolated fractions are 0.3% and 71.1%, respectively.
(D) Starting with immunized BALB/c mouse bone marrow, the plasma cell (CD138+CD267 (TACI)+) content of the isolated fraction is typically 85.5 ± 9.8% (mean ± SD using the purple EasySep? Magnet). In the above example, the purities of the start and isolated fractions are 0.6% and 89.0%, respectively.
(E) Isolated CD138+ cells or total splenocytes from mice immunized with various antigens were fused with Sp2/0 mouse myeloma cells and plated in semi-solid medium using ClonaCell?-HY Hybridoma Kit (Catalog #03800). The % antigen-specific hit rate was determined by ELISA. The % antigen-specific hit rates for total splenocytes and CD138+ cells were 3.2 ± 4.0% and 56.1 ± 37.2% (mean ± SD), respectively.
(F) Isolated CD138+ cells or total bone marrow cells from mice immunized with various antigens were fused with Sp2/0 mouse myeloma cells and plated in semi-solid medium using ClonaCell?-HY Hybridoma Kit (Catalog #03800). The % antigen-specific hit rate was determined by ELISA. The % antigen-specific hit rates for total bone marrow cells and CD138+ cells were 0% and 31.9 ± 46.0% (mean ± SD), respectively.
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EasySep? Magnet
EasySep? Buffer
